Pyrolysis-mass spectrometry has been used to characterize the 17 biological materials including bacteria and proteins. In this study, an in situ thermal-hydrolysis methylation(THM) procedure using tetramethylammonium hydroxide(TMAH) was employed. The biological materials are ionized using chemical ionization(CI) method with ethanol by ion trap mass spectrometer(ITMS), which attached with our own made pyrolyzer module, and then their pyrolysis mass spectra were obtained. The major distinct characteristic peaks were selected from all the range of mass spectra, and analyzed using principal component analysis(PCA) method to assess the classification/identification possibility of biological materials.